title: Direct transformation from amorphous to crystalline calcium phosphate facilitated by motif-programmed artificial proteins.
abstract: An animal's hard tissue is mainly composed of crystalline calcium phosphate. In vitro, small changes in the reaction conditions affect the species of calcium phosphate formed, whereas, in vivo, distinct types of crystalline calcium phosphate are formed in a well-controlled spatiotemporal-dependent manner. A variety of proteins are involved in hard-tissue formation; however, the mechanisms by which they regulate crystal growth are not yet fully understood. Clarification of these mechanisms will not only lead to the development of new therapeutic regimens but will also provide guidance for the application of biomineralization in bionanotechnology. Here, we focused on the peptide motifs present in dentin matrix protein 1 (DMP1), which was previously shown to enhance hydroxylapatite (HAP) formation when immobilized on a glass substrate. We synthesized a set of artificial proteins composed of combinatorial arrangements of these motifs and successfully obtained clones that accelerated formation of HAP without immobilization. Time-resolved static light-scattering analyses revealed that, in the presence of the protein, amorphous calcium phosphate (ACP) particles increased their fractal dimension and molecular mass without increasing their gyration radii during a short period before precipitation. The protein thus facilitated reorganization of the internal structure of amorphous particles into ordered crystalline states, i.e., the direct transformation of ACP to HAP, thereby acting as a nucleus for precipitation of crystalline calcium phosphate. Without the protein, the fractal dimension, molecular mass, and gyration radii of ACP particles increased concurrently, indicating heterogeneous growth transformation.
authors.affiliation: Division of Protein Engineering, Cancer Institute, Japanese Foundation for Cancer Research, Koto, Tokyo 135-8550, Japan.
authors.forename: Toru
authors.initials: T
authors.lastname: Tsuji
authors.forename: Kazuo
authors.initials: K
authors.lastname: Onuma
authors.forename: Akira
authors.initials: A
authors.lastname: Yamamoto
authors.forename: Mayumi
authors.initials: M
authors.lastname: Iijima
authors.forename: Kiyotaka
authors.initials: K
authors.lastname: Shiba
mesh_terms: D020816:Amino Acid Motifs; D001672:Biocompatible Materials / Q000737:chemistry; D002130:Calcium Phosphates / Q000737:chemistry*; D017886:Durapatite / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D046529:Microscopy, Electron, Transmission; D015202:Protein Engineering; D013059:Spectrum Analysis, Raman; D014961:X-Ray Diffraction
chemical_list: D001672:Biocompatible Materials; D002130:Calcium Phosphates; D016326:Extracellular Matrix Proteins; C519480:amorphous calcium phosphate; D017886:Durapatite